Liquid chromatography/photodiode array detection for determination of strychnine in blood: a fatal case report.

A sensitive method for the identification and quantitation of the toxic alkaloids strychnine and brucine from postmortem specimens has been established. After solid-phase extraction using Oasis | MCX cartridges the extracts were analyzed by highperformance liquid chromatography with diode.array detection. The limit of detection was 0.5 ng/mL blood for strychnine and brucine, and the limit of quantitation was 5 ng/mL blood for strychnine and brucine. The method was applied for the analysis of blood and gastric contents of a 34-year-old female who died after ingestion of a packet of herbal medicine powder containing the seeds of Strychnos nux-vomica L. Strychnine and brucine were detected in all the samples. The concentration in our case is consistent with that in previous reports.

Analysis of Strychnine and Brucine in Postmortem Specimens by RP-HPLC: A Case Report of Fatal Intoxication

1. The urinary and faecal excretions of radioactivity, in rats dosed with 3H-strychnine at 0.5 mg/kg subcutaneously, were approx. 30% and 65% of the dose in 7 days, respectively. The radioactivity was mostly excreted within 24 h. 2. Approx. 6% and 3% dose was excreted into urine and faeces, respectively, as unchanged strychnine. 3. Urinary metabolites were extracted from rat urine and purified by silica gel column, t.l.c. and h.p.l.c. Six urinary metabolites, namely, strychnine N-oxide, 21 alpha,22 alpha-dihydroxy-22-hydrostrychnine, 21 alpha,22 beta-dihydroxy-22-hydrostrychnine, 2-hydroxy-strychnine, strychnine 21,22-epoxide and 16-hydroxystrychnine, were identified by comparison with authentic samples by g.l.c.-mass spectrometry. The major metabolites of strychnine in vivo was strychnine 21,22-epoxide.

Metabolic fate of strychnine in rats

A rapid, simple, and sensitive method has been developed for the identification and quantitation of strychnine in human blood. The sample cleanup procedure involved solid-phase extraction with Oasis® HLB cartridges. The extracts were analyzed by gas chromatography-electron impact ionization-mass spectrometry. Limits of detection (LOD) and quantitation (LOQ) were 0.03 and 0.10 μg/mL, respectively, and the method was found to be linear between the LOQ and 2.5 μg/mL, with a correlation coefficient of 0.9994. Intra- and interday precision and accuracy were determined at both low and high concentrations (0.50 and 2.00 μg/mL). The CVs ranged from 5.63 to 8.50% and bias was within ± 10% of the true value. Mean recovery of strychnine was 90.7%. Because of its simplicity and speed, the described method can be applied in forensic toxicology laboratories to determine this alkaloid in whole blood samples. Also, the fact that only 0.5 mL of blood is required to accomplish the analysis make this procedure useful in situations where several exams are needed and the sample volume is limited.

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Determination of strychnine in human blood using solid-phase extraction and GC-EI-MS.

Clinical and therapeutic aspects of non-ketotic hyperglycinaemia

A high performance liquid chromatography (HPLC) method was developed for the quantitation of strychnine in urine of children with nonketotic hyperglycinaemia and other developmental disorders treated with the alkaloid. Mobile and stationary phases were polar, i.e. methanol-water-330 g/kg ammonia (volumes, 85 ml + 14.2 ml + 0.8 ml) and LiChrosorb Si-60, 7 microns. Brucine was the internal standard. Extraction was performed by the Extrelut technique. At strychnine nitrate concentrations in urine of 21, 126, and 70 micrograms/l, recovery was 92.1 +/- 8.7, 98.1 +/- 2.7, and 102.5 +/- 2.7%. A child with nonketotic hyperglycinaemia under continued strychnine treatment excreted 1 to 13.6% of the daily dose unmetabolized in urine. The method was also suitable for the estimation of unreacted strychnine in tissue extracts. The fast disappearance in vitro of strychnine from a guinea pig liver preparation was confirmed.

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W. Steiner

Papers

A New High Performance Liquid Chromatography (HPLC) Method for the Quantitation of Strychnine in Urine and Tissue Extracts